The Vascular Biochemistry and Molecular Biology Core is designed to provide investigators with access to[unreadable] routine as well as state-of-the-art molecular and biochemical tools, expertise, and techniques. The central theme[unreadable] of this Program Project is that exercise induces beneficial phenotypic changes in vascular cells. Our long-term[unreadable] objectives are to advance the understanding of the molecular, biochemical, cellular, and integrative mechanisms[unreadable] that underlie these training effects. The application of molecular and biochemical techniques will be integral in[unreadable] the exploration of the mechanisms underlying the physiological observations made by this PPG group. The[unreadable] Core's activity reflects a centralization and expansion of services that the Core Leader has successfully provided[unreadable] to PPG investigators for the past decade. The Core laboratory is fully equipped and dedicated to address the[unreadable] stated goals and is directed and staffed by well-qualified personnel. The following Aims are based on the needs[unreadable] of the PPG investigators and reflect a compilation of procedures and techniques that will ensure the efficient[unreadable] completion of each PPG project. The long-term goal of this Core is to track the expression and modulation of[unreadable] specific proteins throughout the coronary and peripheral vasculature and to demonstrate an effect of exercise[unreadable] and/or high-fat-cholesterol on these features. Aim 1 is to perform reverse transcriptase-PCR (RT-PCR) to detect[unreadable] specific mRNAs expressed throughout the vasculature. Aim 2 is to perform real-time PCR in order to more[unreadable] accurately quantitate mRNA expression in select samples from the vasculature of control and experimental[unreadable] animals. Aim 3 is to utilize immunoblots and co-immunoprecipitations to detect and relatively quantitate specific[unreadable] proteins and interactions expressed throughout the circulation. Aim 4a is to use in situ PCR to demonstrate[unreadable] tissue-specific expression of select mRNA species. Aim 4b is to use laser-capture microdissection to selectively[unreadable] isolate and recover select cells from sections of vascular tissue. Real-time PCR will then be used to assess[unreadable] mRNA levels in the recovered cells. Aim 5 is to generate two commercial libraries (one from conduit arteries and[unreadable] one from resistance arterioles) and screen for cDNAs encoding select proteins from the coronary vasculature.[unreadable] Aim 6 is to prepare adeno-associated viral constructs for the transduction of various proteins and siRNA into[unreadable] vascular cells. Aim 7 is to coordinate the commercial preparation of select porcine-specific antibodies. Aim 8 is[unreadable] to prepare and analyze peripheral blood cells from transgenic and non-transgenic pigs. Aim 9 is to provide[unreadable] additional standard techniques as well as pioneer new methods as a resource for PPG investigators.